Duration Of Prior Wakefulness Research Articles

Progesterone Might Be an Active Component of the Sleep-Wake Homeostatic Mechanism

Abstract Background: Using a randomized, double blind, placebo-controlled, crossover protocol, we have shown previously that progesterone may prevent sleep disturbances but has no action on undisturbed sleep. In that study, 8 healthy postmenopausal women took daily at 2300 h for 3 wk a capsule of either 300 mg of progesterone or placebo. Sleep was polygraphically recorded during the last two nights and, during the second night, blood samples were collected at 15-min intervals using an iv catheter,. During the first night, sleep was normal under placebo and progesterone had no effect. During the second night, blood sampling procedure was associated under placebo with marked sleep disturbances, which were considerably reduced under progesterone: at the group level, mean ± SEM duration of wake after sleep onset (WASO) dropped from 152 ± 37 min to 71 ± 19 min (P = 0.01), and slow-wave sleep (SWS) duration increased from 53 ± 6 min to 79 ± 10 min (P = 0.04). Objective: To submit individual data collected in that study to new analyses designed to further investigate possible mechanism(s) of progesterone actions on sleep architecture. Results: Among individual subjects, no relation could be evidenced between progesterone levels and sleep variables, or between individual progesterone-associated improvements and absolute values of corresponding sleep variables under placebo. By contrast, for WASO and SWS, significant positive correlations (Spearman test) were evidenced between individual responses to progesterone (i.e. the difference, during night 2, between value under progesterone and value under placebo) and corresponding individual alterations caused under placebo by the blood sampling procedure (i.e. the difference, under placebo, between value during night 1 and value during night 2): WASO: rs = 0.74, P = 0.037, n = 8; SWS: rs = 0.86, P = 0.014, n = 7. (Pearson test yielded similar results: WASO: r = 0.85, P = 0.008; SWS: r = 0.76, P = 0.047). Conclusions: Although they obviously need to be confirmed by larger studies performed in a variety of clinical conditions, the present findings suggest that progesterone action on sleep architecture is specifically tailored to restore individual normality rather than group normality. Since SWS is mainly regulated by the sleep-wake homeostatic mechanism relating sleep pressure to the duration of prior wakefulness, it is tempting to speculate that progesterone might be an active component of this mechanism.

Evidence That Homeostatic Sleep Regulation Depends on Ambient Lighting Conditions during Wakefulness.

We examined whether ambient lighting conditions during extended wakefulness modulate the homeostatic response to sleep loss as indexed by. slow wave sleep (SWS) and electroencephalographic (EEG) slow-wave activity (SWA) in healthy young and older volunteers. Thirty-eight young and older participants underwent 40 hours of extended wakefulness [i.e., sleep deprivation (SD)] once under dim light (DL: 8 lux, 2800 K), and once under either white light (WL: 250 lux, 2800 K) or blue-enriched white light (BL: 250 lux, 9000 K) exposure. Subjective sleepiness was assessed hourly and polysomnography was quantified during the baseline night prior to the 40-h SD and during the subsequent recovery night. Both the young and older participants responded with a higher homeostatic sleep response to 40-h SD after WL and BL than after DL. This was indexed by a significantly faster intra-night accumulation of SWS and a significantly higher response in relative EEG SWA during the recovery night after WL and BL than after DL for both age groups. No significant differences were observed between the WL and BL condition for these two particular SWS and SWA measures. Subjective sleepiness ratings during the 40-h SD were significantly reduced under both WL and BL compared to DL, but were not significantly associated with markers of sleep homeostasis in both age groups. Our data indicate that not only the duration of prior wakefulness, but also the experienced illuminance during wakefulness affects homeostatic sleep regulation in humans. Thus, working extended hours under low illuminance may negatively impact subsequent sleep intensity in humans.

The alerting effect of the wake maintenance zone during 40 hours of sleep deprivation

Under entrained conditions, the accumulation of homeostatic sleep pressure in the evening is opposed by a strong circadian arousal signal prior to the dim light melatonin onset, called the Wake Maintenance Zone (WMZ). This study aimed at investigating the impact of the WMZ on different cognitive performance tests, as well as on subjective and objective sleepiness. Twelve young male participants completed a constant routine protocol with 40 h of extended wakefulness that included two WMZs. Cognitive tests and saliva samples were assessed hourly, while the electroencephalogram (EEG) was recorded continuously. Participants improved in cognitive response inhibition during WMZ1 (13.5 h awake) and sustained attention during WMZ2 (37.5 h awake), but not in higher executive function tests. There were significant EEG power density reductions in the delta/theta frequency range during WMZ1 and in delta/theta, alpha, and sigma/beta ranges during WMZ2, with a greater change in the sigma/beta range during WMZ2 compared to WMZ1. EEG power reductions coincided during WMZ1 with stable subjective sleepiness and sustained attention. During WMZ2, EEG power reductions were more pronounced and coincided with improved sustained attention. Our results suggest the circadian arousal signal in the evening differently modulates cognitive functions and EEG power depending on the duration of prior wakefulness.

Molecular Mechanisms of Sleep Homeostasis in Flies and Mammals.

Sleep is homeostatically regulated with sleep pressure accumulating with the increasing duration of prior wakefulness. Yet, a clear understanding of the molecular components of the homeostat, as well as the molecular and cellular processes they sense and control to regulate sleep intensity and duration, remain a mystery. Here, we will discuss the cellular and molecular basis of sleep homeostasis, first focusing on the best homeostatic sleep marker in vertebrates, slow wave activity; second, moving to the molecular genetic analysis of sleep homeostasis in the fruit fly Drosophila; and, finally, discussing more systemic aspects of sleep homeostasis.

A mechanism for sickness sleep: lessons from invertebrates

During health, animal sleep is regulated by an internal clock and by the duration of prior wakefulness. During sickness, sleep is regulated by cytokines released from either peripheral cells or from cells within the nervous system. These cytokines regulate central nervous system neurons to induce sleep. Recent research in the invertebrates Caenorhabditis elegans and Drosophila melanogaster has led to new insights into the mechanism of sleep during sickness. Sickness is triggered by exposure to environments such as infection, heat, or ultraviolet light irradiation, all of which cause cellular stress. Epidermal growth factor is released from stressed cells and signals to activate central neuroendocrine cell(s). These neuron(s) release neuropeptides including those containing an amidated arginine(R)-phenylalanine(F) motif at their C-termini (RFamide peptides). Importantly, mechanisms regulating sickness sleep are partially distinct from those regulating healthy sleep. We will here review key findings that have elucidated the central neuroendocrine mechanism of sleep during sickness. Adaptive mechanisms employed in the control of sickness sleep may play a role in correcting cellular homeostasis after various insults. We speculate that these mechanisms may play a maladaptive role in human pathological conditions such as in the fatigue and anorexia associated with autoimmune diseases, with major depression, and with unexplained chronic fatigue.

Homeostatic sleep pressure is the primary factor for activation of cortical nNOS/NK1 neurons.

Cortical interneurons, immunoreactive for neuronal nitric oxide synthase (nNOS) and the receptor NK1, express the functional activity marker Fos selectively during sleep. NREM sleep 'pressure' is hypothesized to accumulate during waking and to dissipate during sleep. We reported previously that the proportion of Fos(+) cortical nNOS/NK1 neurons is correlated with established electrophysiological markers of sleep pressure. As these markers covary with the amount of NREM sleep, it remained unclear whether cortical nNOS/NK1 neurons are activated to the same degree throughout NREM sleep or whether the extent of their activation is related to the sleep pressure that accrued during the prior waking period. To distinguish between these possibilities, we used hypnotic medications to control the amount of NREM sleep in rats while we varied prior wake duration and the resultant sleep pressure. Drug administration was preceded by 6 h of sleep deprivation (SD) ('high sleep pressure') or undisturbed conditions ('low sleep pressure'). We find that the proportion of Fos(+) cortical nNOS/NK1 neurons was minimal when sleep pressure was low, irrespective of the amount of time spent in NREM sleep. In contrast, a large proportion of cortical nNOS/NK1 neurons was Fos(+) when an equivalent amount of sleep was preceded by SD. We conclude that, although sleep is necessary for cortical nNOS/NK1 neuron activation, the proportion of cells activated is dependent upon prior wake duration.

Neurobiologic Mechanisms of Sleep and Wakefulness

Neurobiologic Mechanisms of Sleep and Wakefulness

Low-frequency rhythmic electrocutaneous hand stimulation during slow-wave night sleep: Physiological and therapeutic effects

Neocortical EEG slow wave activity (SWA) in the delta frequency band (0.5-4.0 Hz) is a hallmark of slow wave sleep (SWS) and its power is a function of prior wake duration and an indicator of a sleep need. SWS is considered the most important stage for realization of recovery functions of sleep. Possibility of impact on characteristics of a night sleep by rhythmic (0.8-1.2 Hz) subthreshold electrocutaneous stimulation of a hand during SWS is shown: 1st night--adaptation, 2nd night--control, 3d and 4th nights--with stimulation during SWA stages ofa SWS. Stimulation caused significant increase in average duration of SWS and EEG SWA power (in 11 of 16 subjects), and also well-being and mood improvement in subjects with lowered emotional tone. It is supposed that the received result is caused by functioning of a hypothetical mechanism directed on maintenance and deepening of SWS and counteracting activating, awakening influences of the afferent stimulation. The results can be of value both for understanding the physiological mechanisms of sleep homeostasis and for development of non-pharmacological therapy of sleep disorders.

Cul3 and the BTB Adaptor Insomniac Are Key Regulators of Sleep Homeostasis and a Dopamine Arousal Pathway in Drosophila

Sleep is homeostatically regulated, such that sleep drive reflects the duration of prior wakefulness. However, despite the discovery of genes important for sleep, a coherent molecular model for sleep homeostasis has yet to emerge. To better understand the function and regulation of sleep, we employed a reverse-genetics approach in Drosophila. An insertion in the BTB domain protein CG32810/insomniac (inc) exhibited one of the strongest baseline sleep phenotypes thus far observed, a ∼10 h sleep reduction. Importantly, this is coupled to a reduced homeostatic response to sleep deprivation, consistent with a disrupted sleep homeostat. Knockdown of the INC-interacting protein, the E3 ubiquitin ligase Cul3, results in reduced sleep duration, consolidation, and homeostasis, suggesting an important role for protein turnover in mediating INC effects. Interestingly, inc and Cul3 expression in post-mitotic neurons during development contributes to their adult sleep functions. Similar to flies with increased dopaminergic signaling, loss of inc and Cul3 result in hyper-arousability to a mechanical stimulus in adult flies. Furthermore, the inc sleep duration phenotype can be rescued by pharmacological inhibition of tyrosine hydroxylase, the rate-limiting enzyme for dopamine biosynthesis. Taken together, these results establish inc and Cul3 as important new players in setting the sleep homeostat and a dopaminergic arousal pathway in Drosophila.

Sleep, Wake and Phase Dependent Changes in Neurobehavioral Function under Forced Desynchrony

The homeostatic-circadian regulation of neurobehavioral functioning is not well understood in that the role of sleep dose in relation to prior wake and circadian phase remains largely unexplored. The aim of the present study was to examine the neurobehavioral impact of sleep dose at different combinations of prior wake and circadian phase. A between-participant design involving 2 forced desynchrony protocols varying in sleep dose. Both protocols comprised 7 repetitions of a 28-h sleep/wake cycle. The sleep dose in a standard protocol was 9.33 h per 28-h day and 4.67 h in a sleep-restricted protocol. A time-isolation laboratory at the Centre for Sleep Research, the University of South Australia. A total of 27 young healthy males participated in the study with 13 in the standard protocol (age 22.5 ± 2.2 y) and 14 in the sleep-restricted protocol (age 21.8 ± 3.8 y). Wake periods during both protocols were approximately 4 h delayed each 28-h day relative to the circadian system, allowing performance testing at different combinations of prior wake and circadian phase. The manipulation in sleep dose between the 2 protocols, therefore, allowed the impact of sleep dose on neurobehavioral performance to be examined at various combinations of prior wake and circadian phase. Neurobehavioral function was assessed using the psychomotor vigilance task (PVT). There was a sleep dose × circadian phase interaction effect on PVT performance such that sleep restriction resulted in slower and more variable response times, predominantly during the biological night. This interaction was not altered by prior wakefulness, as indicated by a nonsignificant sleep dose × circadian phase × prior wake interaction. The performance consequence of sleep restriction in our study was prominent during the biological night, even when the prior wake duration was short, and this performance consequence was in forms of waking state instability. This result is likely due to acute homeostatic sleep pressure remaining high despite the sleep episode.

Functional Anatomy of Non-REM Sleep.

The state of non-REM sleep (NREM), or slow wave sleep, is associated with a synchronized EEG pattern in which sleep spindles and/or K complexes and high-voltage slow wave activity (SWA) can be recorded over the entire cortical surface. In humans, NREM is subdivided into stages 2 and 3–4 (presently named N3) depending on the proportions of each of these polygraphic events. NREM is necessary for normal physical and intellectual performance and behavior. An overview of the brain structures involved in NREM generation shows that the thalamus and the cerebral cortex are absolutely necessary for the most significant bioelectric and behavioral events of NREM to be expressed; other structures like the basal forebrain, anterior hypothalamus, cerebellum, caudal brain stem, spinal cord and peripheral nerves contribute to NREM regulation and modulation. In NREM stage 2, sustained hyperpolarized membrane potential levels resulting from interaction between thalamic reticular and projection neurons gives rise to spindle oscillations in the membrane potential; the initiation and termination of individual spindle sequences depends on corticothalamic activities. Cortical and thalamic mechanisms are also involved in the generation of EEG delta SWA that appears in deep stage 3–4 (N3) NREM; the cortex has classically been considered to be the structure that generates this activity, but delta oscillations can also be generated in thalamocortical neurons. NREM is probably necessary to normalize synapses to a sustainable basal condition that can ensure cellular homeostasis. Sleep homeostasis depends not only on the duration of prior wakefulness but also on its intensity, and sleep need increases when wakefulness is associated with learning. NREM seems to ensure cell homeostasis by reducing the number of synaptic connections to a basic level; based on simple energy demands, cerebral energy economizing during NREM sleep is one of the prevalent hypotheses to explain NREM homeostasis.

THE INFLUENCE OF CIRCADIAN PHASE AND PRIOR WAKE ON NEUROMUSCULAR FUNCTION

Previous forced desynchrony (FD) studies have shown that neurobehavioral function is affected by circadian phase and duration of prior wakefulness. There is some evidence that neuromuscular function may also be affected by circadian phase and prior wake, but these effects have not been systematically investigated. This study examined the effects of circadian phase and prior wake on two measures of neuromuscular function—postural balance (PB) and maximal grip strength (MGS)—using a 28-h FD protocol. Eleven male participants (mean ± SD: 22.7 ± 2.5 yr) lived in a sound-attenuated, light- and temperature-controlled time-isolation laboratory for 12 days. Following two training days and a baseline day, participants were scheduled to seven 28-h FD days, with the ratio between sleep opportunity and wake spans kept constant (i.e., 9.3 h sleep period and 18.7 h wake period). PB was measured during 1 min of quiet standing on a force platform. MGS of the dominant hand was measured using a dynamometer. These two measures were obtained every 2.5 h during wake. Core body temperature was continuously recorded with rectal thermistors to determine circadian phase. For both measures of neuromuscular function, individual data points were assigned a circadian phase and a level of prior wake. Data were analyzed by repeated-measures analysis of variance (ANOVA) with two within-subjects factors: circadian phase (six phases) and prior wake (seven levels). For MGS, there was a main effect of circadian phase, but no main effect of prior wake. For PB, there were no main effects of circadian phase or prior wake. There were no interactions between circadian phase and prior wake for MGS or PB. The significant effect of circadian phase on muscle strength is in agreement with previous reports in the literature. In terms of prior wake, both MGS and PB remained relatively stable across wake periods, indicating that neuromuscular function may be more robust than neurobehavioral function when the duration of wakefulness is within a normal range (i.e., 18.7 h). (Author correspondence: charli.sargent@unisa.edu.au)

Homeostatic regulation of sleep in the white-crowned sparrow (Zonotrichia leucophrys gambelii)

BackgroundSleep is regulated by both a circadian and a homeostatic process. The homeostatic process reflects the duration of prior wakefulness: the longer one stays awake, the longer and/or more intense is subsequent sleep. In mammals, the best marker of the homeostatic sleep drive is slow wave activity (SWA), the electroencephalographic (EEG) power spectrum in the 0.5–4 Hz frequency range during non-rapid eye movement (NREM) sleep. In mammals, NREM sleep SWA is high at sleep onset, when sleep pressure is high, and decreases progressively to reach low levels in late sleep. Moreover, SWA increases further with sleep deprivation, when sleep also becomes less fragmented (the duration of sleep episodes increases, and the number of brief awakenings decreases). Although avian and mammalian sleep share several features, the evidence of a clear homeostatic response to sleep loss has been conflicting in the few avian species studied so far. The aim of the current study was therefore to ascertain whether established markers of sleep homeostasis in mammals are also present in the white-crowned sparrow (Zonotrichia leucophrys gambelii), a migratory songbird of the order Passeriformes. To accomplish this goal, we investigated amount of sleep, sleep time course, and measures of sleep intensity in 6 birds during baseline sleep and during recovery sleep following 6 hours of sleep deprivation.ResultsContinuous (24 hours) EEG and video recordings were used to measure baseline sleep and recovery sleep following short-term sleep deprivation. Sleep stages were scored visually based on 4-sec epochs. EEG power spectra (0.5–25 Hz) were calculated on consecutive 4-sec epochs. Four vigilance states were reliably distinguished based on behavior, visual inspection of the EEG, and spectral EEG analysis: Wakefulness (W), Drowsiness (D), slow wave sleep (SWS) and rapid-eye movement (REM) sleep. During baseline, SWA during D, SWS, and NREM sleep (defined as D and SWS combined) was highest at the beginning of the major sleep period and declined thereafter. Moreover, peak SWA in both SWS and NREM sleep increased significantly immediately following sleep deprivation relative to baseline.ConclusionAs in mammals, sleep deprivation in the white-crowned sparrow increases the intensity of sleep as measured by SWA.

Sleep homeostasis in the rat in the light and dark period

Sleep is regulated by the interaction of a homeostatic (Process S) and a circadian component. The duration of prior wakefulness is the main factor influencing subsequent sleep duration and its intensity. We investigated in the rat whether the sleep–wake history before sleep deprivation (SD) contributes to the effects of sleep loss incurred during the SD. A 24-h baseline recording was followed by 6 h SD at light onset (SD-Light, n = 7), or at dark onset (SD-Dark, n = 8) and 18 h recovery. Both SDs led to a pronounced increase in slow wave activity (SWA, EEG power between 0.75 and 4.0 Hz) in NREM sleep and increased sleep consolidation. The prolongation of sleep episodes was associated with increased intra-episode SWA. The amount of waking before the SD correlated positively with the SWA increase during recovery, and SWA levels before SD were negatively correlated with their subsequent increase. The time-course of SWA (Process S) as well as of single frequency bins within the SWA band was successfully simulated based on vigilance-state distribution. The time constant of the exponential monotonic decay ( T d) was higher for the 0.75–1.0 Hz bin compared to all remaining frequency bins of the SWA band, reflecting a slower process determining the slow EEG component during sleep. The data show that the homeostatic response after SD, consisting of increased sleep intensity and sleep consolidation is determined by a combination of SD and the preceding vigilance-state history. The slower dynamics of low frequency delta power compared to fast delta frequencies point to heterogeneity within the traditionally defined SWA band.

Exploratory Behavior, Cortical BDNF Expression, and Sleep Homeostasis

Slow-wave activity (SWA; 0.5-4.0 Hz) during non-rapid eye movement (NREM) sleep is a reliable indicator of sleep need, as it increases with the duration of prior wakefulness and decreases during sleep. However, which biologic process occurring during wakefulness is responsible for the increase of sleep SWA remains unknown. The aim of the study was to determine whether neuronal plasticity underlies the link between waking activities and the SWA response. We manipulated, in rats, the amount of exploratory activity while maintaining the total duration of waking constant. We then measured the extent to which exploration increases cortical expression of plasticity-related genes (BDNF, Arc, Homer, NGFI-A), and the SWA response once the animals were allowed to sleep. Basic neurophysiology and molecular laboratory. Male Wistar Kyoto rats (250-300 g; 2-3 month old). None. We found that, within the same animal, the amount of exploratory behavior during wakefulness could predict the extent to which BDNF was induced, as well as the extent of the homeostatic SWA response during subsequent sleep. This study suggests a direct link between the synaptic plasticity triggered by waking activities and the homeostatic sleep response and identifies BDNF as a major mediator of this link at the molecular level.

Gene expression in the rat brain during sleep deprivation and recovery sleep: an Affymetrix GeneChip® study

Previous studies have demonstrated that macromolecular synthesis in the brain is modulated in association with the occurrence of sleep and wakefulness. Similarly, the spectral composition of electroencephalographic activity that occurs during sleep is dependent on the duration of prior wakefulness. Since this homeostatic relationship between wake and sleep is highly conserved across mammalian species, genes that are truly involved in the electroencephalographic response to sleep deprivation might be expected to be conserved across mammalian species. Therefore, in the rat cerebral cortex, we have studied the effects of sleep deprivation on the expression of immediate early gene and heat shock protein mRNAs previously shown to be upregulated in the mouse brain in sleep deprivation and in recovery sleep after sleep deprivation. We find that the molecular response to sleep deprivation and recovery sleep in the brain is highly conserved between these two mammalian species, at least in terms of expression of immediate early gene and heat shock protein family members. Using Affymetrix Neurobiology U34 GeneChips® , we also screened the rat cerebral cortex, basal forebrain, and hypothalamus for other genes whose expression may be modulated by sleep deprivation or recovery sleep. We find that the response of the basal forebrain to sleep deprivation is more similar to that of the cerebral cortex than to the hypothalamus. Together, these results suggest that sleep-dependent changes in gene expression in the cerebral cortex are similar across rodent species and therefore may underlie sleep history-dependent changes in sleep electroencephalographic activity.

No interference of task complexity with circadian rhythmicity in a constant routine protocol

Time course in task performance has been studied extensively. In particular, the investigation of circadian rhythmicity in task performance that varied in complexity. However, these studies disclosed heterogeneous outcomes. This could be the result of confounding exogenous factors, the use of diverse tasks, as well as accumulating sleepiness interfering with the underlying circadian drive. The present study varied task demands systematically within a single task and a dual task, using a constant routine protocol to examine the unmasked influence of the endogenous circadian oscillator on the periodicity of performance. Moreover, the subjects were divided into an early-start and a late-start group to estimate the potential interaction of circadian rhythmicity with the duration of prior wakefulness. The results revealed a distinct congruence in the circadian rhythms of all performance measures, with which prior wakefulness (< 40 h) did not interact. Also, single-task as well as dual-task complexity did not interfere with circadian rhythmicity. In conclusion, when sufficiently controlled for masking exogenous factors, task complexity is removed from the underlying circadian rhythms.

A social conflict increases EEG slow-wave activity during subsequent sleep

Electroencephalogram (EEG) slow-wave activity (SWA) during non-rapid eye movement (NREM) sleep is widely viewed as an indicator of sleep debt and sleep intensity. In a previous study, we reported a strong increase in SWA during NREM sleep after a social conflict in rats. To test whether this increase in SWA reflects normal physiological sleep or an unrelated by-product of the stress, we now measured the effect of a conflict in combination with extended sleep deprivation by means of gentle handling. We anticipated that if the social defeat-induced SWA reflects a true sleep debt, the drive for it would persist during the extended wakefulness. Male rats were subjected to a 1-h social conflict followed by 5 h of sleep deprivation by gentle handling or to 6 h gentle handling alone. The manipulations took place during the second half of the dark phase and recovery sleep was recorded during the subsequent light phase. Neither of the two procedures caused a significant change in the total duration of NREM or REM sleep thereafter. Yet, both modes of sleep deprivation induced a strong increase in SWA during NREM sleep. This SWA was significantly higher for 6 h after sleep deprivation consisting of a social conflict followed by gentle handling, as compared to sleep deprivation by handling alone. Thus, the SWA increasing effect of the conflict persisted during the extended wakefulness. The data confirm that social defeat stress accelerates the build up of sleep debt and support the notion that sleep debt and subsequent NREM sleep intensity not only depend on the duration of prior wakefulness but also on what animals experience during that waking.

The Homeostatic Regulation of Sleep Need Is under Genetic Control

Delta power, a measure of EEG activity in the 1-4 Hz range, in slow-wave sleep (SWS) is in a quantitative and predictive relationship with prior wakefulness. Thus, sleep loss evokes a proportional increase in delta power, and excess sleep a decrease. Therefore, delta power is thought to reflect SWS need and its underlying homeostatically regulated recovery process. The neurophysiological substrate of this process is unknown and forward genetics might help elucidate the nature of what is depleted during wakefulness and recovered during SWS. We applied a mathematical method that quantifies the relationship between the sleep-wake distribution and delta power to sleep data of six inbred mouse strains. The results demonstrated that the rate at which SWS need accumulated varied greatly with genotype. This conclusion was confirmed in a "dose-response" study of sleep loss and changes in delta power; delta power strongly depended on both the duration of prior wakefulness and genotype. We followed the segregation of the rebound of delta power after sleep deprivation in 25 BXD recombinant inbred strains by quantitative trait loci (QTL) analysis. One "significant" QTL was identified on chromosome 13 that accounted for 49% of the genetic variance in this trait. Interestingly, the rate at which SWS need decreases did not vary with genotype in any of the 31 inbred strains studied. These results demonstrate, for the first time, that the increase of SWS need is under a strong genetic control, and they provide a basis for identifying genes underlying SWS homeostasis.

Sleep deprivation-induced c-fos and junB expression in the rat brain: effects of duration and timing

Expression of the immediate-early genes (IEGs) c-fos and junB in the rat brain was studied in response to sleep deprivation (SD) starting at four time points during the light phase of a 12:12 light:dark cycle. Animals were confined to slowly rotating wheels for 3 or 6 h in order to prevent sleep. The numbers of c-Fos- and JunB-immunoreactive cells were assessed in seven brain regions previously reported to respond to SD with increased c-fos expression (medial preoptic area (MPA), cortex, anterior and posterior paraventricular thalamic nuclei, amygdala, caudate-putamen, and laterodorsal tegmental nucleus). While c-Fos was induced by SD in all regions studied, there were differences in levels of induction depending on the duration of deprivation and on the timing of the deprivation period during the light phase. The most robust induction occurred in most regions in response to 3-h deprivation periods beginning 3 h into the light phase. A similarly timed peak of induction was observed in the MPA and cortex after 6 h of SD. In two regions, the posterior paraventricular thalamic nucleus and amygdala, 6 h of deprivation induced greater c-Fos immunoreactivity than did 3 h of deprivation, collapsed across all phases tested. Increased JunB immunoreactivity in response to either duration of deprivation was more limited and was significant only in the MPA, cortex, caudate-putamen and amygdala. c-Fos and JunB immunoreactivity in the paraventricular hypothalamic nucleus was low and similar in control and deprived animals. These results indicate that both duration of prior wakefulness and time of day influence the extent of IEG expression differentially in brain regions responsive to SD. The results also suggest that the posterior paraventricular thalamic nucleus and amygdala might be primarily responsive to length of wakefulness (sleep drive), while the MPA and anterior paraventricular thalamic nucleus might integrate input related to both homeostatic sleep drive and circadian clock influences on sleep regulation.